CN1036398C - Therapeutic nucleosides - Google Patents

Abstract

The present invention provides a compound of formula or a physiologically functional derivative thereof:

wherein R is¹Representing n-propoxy, cyclobutoxy, cyclopropylamino, piperidino or pyrrolidinyl, and further processes for the preparation of, compositions containing and the use of, compounds of formula as antiviral agents.

Description

Therapeutic nucleosides

The present invention relates to new 2 ', 3 '-dideoxy-3 '-the fluoropurine nucleoside compound, and salt, ester and derivative with physiological function, their preparation method contains their pharmaceutical preparation and their application in treatment, is particularly useful for treatment or infection prevention venereal disease poison.

In antiviral chemotherapy field, the minority medicine demonstrates and effectively kills virus activity itself, and because the difficulty of attack virus, host cells infected is not injury-free and stay.Confirm: from a kind of life-cycle processes that changes to another kind, some stage is determined by virus itself in virus.These stages can prove that they are being different from the local vulnerable of any corresponding host cell function fully.Yet because the very big similarity between virus and the host's function, effectively treatment has proved and has been difficult to distinguish them.

The viral pathogen that one class worldwide has great effect is hepatitis virus, particularly hepatitis B virus (HBV).

HBV sees Asian countries usually, and area, nearly the Sahara is popular in Africa.This virus is relevant with the initial stage hepatocellular carcinoma cause of disease, and is considered to cause 80% whole world liver cancer.In the U.S., 10,000 people of surpassing are arranged because of the hospital care of HBV disease every year, on average there are 250 people to die from sudden illness.Estimate that the U.S. has 500,000 to 1,000,000 infectivity cause of disease carrier at present.Surpass 25% pathogen carrier and develop into chronic active hepatitis, and change liver cirrhosis into usually.The annual according to estimates U.S. has 5000 people to die from the liver cirrhosis relevant with HBV approximately, and 1000 people die from the liver cancer relevant with HBV.Both made and used the HBV vaccine in the whole world, still needed effective anti-HBV compound.A large amount of obstinate infected pathogen carriers (estimating that the whole world has 22,000 ten thousand) will can not treated by inoculation, and still have very big danger to be induced into hepatopathy by HBV.These pathogen carriers are contagium of keeping the susceptible individual of disease generation, especially in the endemy area, or high-risk populations, as drug abuser and homosexual.Thereby, still greatly need potent antiviral agent, be used to control chronic infection and the progress that reduces to hepatocellular carcinoma.

Scope by the infectious clinical effect of HBV comprises headache, heating, the uncomfortable ， Evil heart, vomiting, apocleisis and stomachache.Duplicating usually of this virus controlled by immunne response, and recuperation lasting several weeks or several months in human body, may be more serious but infect, can cause aforesaid long-term chronic hepatopathy.

HBV is the small virus that contains DNA that infects human body.For with a member of the kind of the closely related virus of known hepad-naviruses, various viruses infect Mammalia or bird host selectively, as marmot and duck.

76 and 77 the chapters detailed infectious hepatitis of having described, the especially etiology of infectivity HBV of " Fields Virology " (Volume2, Ed., Fields etc. (1990) Raven Press, New York).

Following formula (I) compound has dropped in the scope of the disclosed compound of European patent specification No.031728, but does not have specifically to disclose formula (I) compound or their uses in medical treatment.

Amazing and be unexpectedly, we find that now following formula (I) compound is suitable for treatment or prevention hepatitis virus infection, especially HBV.The present invention at first provides formula (I) compound: Wherein R ' represents positive propoxy, cyclobutoxy group, cyclopropylamino, piperidino-(1-position only), or pyrrolidyl; Or the salt of formula (I) compound, ester or derivative, or any their solvate with physiological function.

Formula (I) compound can be named as follows:

(1) .2-amino-9-(2, the 3-dideoxy-red moss of 3-fluoro-beta-D-

-furan pentose base)-6-positive propoxy-9H-purine;

(2) .2-amino-6-cyclobutoxy group-9-(2,3-dideoxy-3-fluorine

The red moss of-β-D--furan pentose base)-the 9H-purine;

(3) .2-amino-6-(cyclopropylamino)-9-(2,3-dideoxy-3

The red moss of-fluoro-beta-D--furan pentose base)-the 9H-purine;

(4) .2-amino-6-(1-piperidino-(1-position only))-9-(2, the 3-dideoxy

The red moss of-3-fluoro-beta-D--furan pentose base)-the 9H-purine; With

(5) .2-amino-6-(1-pyrrolidyl)-9-(2, the 3-dideoxy

The red moss of-3-fluoro-beta-D--furan pentose base)-the 9H-purine.

Particularly preferred formula (I) compound is 2-amino-6-(cyclopropylamino)-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine and 2-amino-6-(1-pyrrolidyl)-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine.The HBV that these compounds are particularly useful for anti-animal infects, and described animal comprises the people, marmot and duck.

Here used predicate " Piperidino " and Pyrrolidino respectively refer to substituted piperidyl and pyrrolidyl on 1.

Above-mentioned formula (I) compound and its are referred to as The compounds of this invention below having the derivative of physiological function

The present invention also provides the The compounds of this invention that is used for the treatment of more properly to say to be the The compounds of this invention as antiviral agent, for example, is used for the treatment of or the infection prevention hepatitis, as infectivity HBV.

The present invention further comprises:

A). a kind ofly treat or prevent infected animal, for example Mammals comprises people's the viral infectious symptom or the method for effect, comprises with the The compounds of this invention of treatment significant quantity handling described animal.According to the particular content of this aspect of the present invention, infective virus is infectivity HBV.

B). a kind of prevention animal, for example Mammals comprises people's the infectious method of viral infection, particularly HBV, comprises with the The compounds of this invention of treatment significant quantity handling described animal.

C). utilize The compounds of this invention preparation to be used for the treatment of or prevent the infectious medicament of viral infection, especially HBV.

Predicate used herein " derivative with physiological function " is meant any physiologically acceptable salt of formula (I) compound, ester, or the salt of these esters, or to acceptor give with after the compound of (directly or indirectly) such compound or the metabolite of its antiviral activity or resistates can be provided.For example, within the scope of the present invention, the OH base on 5 ' is passed through potential hydrolysable group such as acyl group or alkyl replacement H.

Preferably be included in the ester that the present invention has formula (I) compound in the derivative scope of weight function and comprise carboxylicesters, wherein the non-carbonyl moiety of the carbonyl moiety of ester group is selected from straight or branched alkyl (for example, methyl, n-propyl, the tertiary butyl, or normal-butyl), cycloalkyl, alkoxyalkyl (for example, methoxyl methyl), aralkyl (for example benzyl), aryloxy alkyl (for example, benzyloxy), aryl (phenyl for example, can be by for example halogen, C _1-4Alkyl, or C _1-4Alkoxyl group replaces arbitrarily) or amino); Sulphonate is as alkyl-or aralkyl alkylsulfonyl (for example methylsulfonyl); Amino acid ester (for example, L-valyl or L-isoleucyl-); And single-, two-, or three-phosphoric acid ester.In these esters, except as otherwise noted, described moieties preferably contains 1 to 18 carbon atom, is preferably 1 to 6 carbon atom, more preferably 1 to 4 carbon atom.Any cycloalkyl moiety described in these esters preferably contains 3 to 6 carbon atoms.Any aryl moiety in these esters preferably includes the phenyl that is optionally substituted.Any explanation of any above-claimed cpd also comprises the explanation to its physiologically acceptable salt.

The physiologically acceptable salt of formula (I) compound and the example of its physiologically acceptable derivative comprise the salt that is derived by suitable alkali, as basic metal (for example sodium), and alkaline-earth metal (for example magnesium), ammonium and NX ₄ ⁺(wherein X represents C _1-4Alkyl) salt.Hydrogen atom or amino physiologically acceptable salt comprise organic carboxylate, as acetate, lactic acid salt, tartrate, maleate, isethionate and amber salt: organic sulfonate such as mesylate, esilate, benzene sulfonate and right-tosylate and inorganic acid salt such as hydrogen chlorate, vitriol, phosphoric acid salt and sulfamate.The physiologically acceptable salt of compound with hydroxyl is by the negatively charged ion of described compound and suitable positively charged ion such as Na ⁺, NH ₄ ⁺And NX ₄ ⁺(wherein X represents C _1-4Alkyl) chemical combination and forming.

Use for treatment, the salt of formula (I) compound should be physiologically acceptable, and promptly they are the salt of being derived and being obtained by physiologically acceptable acid or alkali.But the salt of non-physiologically acceptable acid or alkali also can find purposes, for example, is used for preparation or the physiologically acceptable compound of purifying.All salt (obtaining no matter whether derive from physiologically acceptable acid or alkali) include within the scope of the present invention.

The compounds of this invention can be separately or is combined with other therapeutical agent and to be used for the treatment of above-mentioned infection or symptom.Combined treatment of the present invention comprises to at least a formula (I) compound or its physiological function derivative and at least a other pharmaceutical active composition.Activeconstituents or pharmacologically active agent can be taken jointly above or respectively, and when taking respectively, can be simultaneously to using or giving usefulness in order with any order.

To select for the activeconstituents of usefulness and the amount and the selection of correlation time of pharmacologically active agent according to reaching desired combined treatment effect.Preferred combined treatment comprises to having derivative and a kind of following promoting agent of physiological function with a kind of formula (I) compound or its.

This class is treated agent to the effectively further treatment of treatment HBV infection and is comprised Carbovir, Oxathiolan nucleoside analog such as cis-1-(2-methylol)-1,3-oxathiane-5-yl)-cytosine(Cyt) (Cis-1-(2-hydroxymethyl)-1,3-Oxathiolan-5-yl)-Cytosin) or cis-1-(2-methylol)-1,3-oxathiane-5-yl)-5-flurocytosine, 2 ', 3 '-dideoxy-5-ethynyl-3 '-floxuridine, 5-chloro-2 ', 3 '-dideoxy-3 '-floxuridine, 1-(β-D-arbinofuranose base)-5-proyl uridylic, acycloguanosine and Interferon, rabbit are as alpha-interferon.

More preferably combined treatment comprises common giving with a kind of above-mentioned medicament and a kind of formula (I) compound of special name here.

The present invention also further provides the pharmaceutical preparation of The compounds of this invention (being also referred to as activeconstituents here), preparation is given by any suitable route that is fit to the clinical disease for the treatment of and needing to be used for the Mammals of treatment to comprise people's (" acceptor "), suitable route comprises oral, rectal administration, nasal administration, surperficial administration (comprises cheek, hypogloeeis and percutaneous drug delivery), vagina administration and parenterai administration (comprise subcutaneous, intramuscular, intravenously and intradermal administration).Be not difficult to know that preferred approach changes with following condition: receptor's body weight, age and sex, the character of infection and selected activeconstituents.

Treat viral infection, comprise that the amount that HBV infects required The compounds of this invention depends on series of factors, comprise sanatory severity of institute and receptor's physical examination, and finally determine by nursing doctor's processing.

Usually, to viral infection, as the scope of the optimal dose of the treatment of HBV be receptor every day every kg body weight 0.5 to 120mg, preferable range be every day every kg body weight 1 to 90mg, most preferred range for whenever in every kg body weight 2 to 60mg.Optimal dose is the about 10mg of every kg body weight every day.Except as otherwise noted, the weight of all activeconstituentss is calculated by formula (1) compound parent.For the physiologically acceptable salt of formula (I) compound, ester or other have derivative or any its solvate of physiological function, and quantity increases in proportion.Required dosage preferably at twice, three times, four times, five times, six administrations, or multiple low dose administration under suitable timed interval in a day.These low doses can be given usefulness with unit dosage form, and for example, the per unit dosage form contains 1 to 1500mg, is preferably 5 to 1000mg, most preferably is 10 to 700mg activeconstituentss.Perhaps, according to the needs of receptor's illness, dosage can be given by the continuous infusion mode and use.

In theory, activeconstituents should be given to use active compound peak plasma concentration about 0.25 to about 100 μ n, is preferably about 0.5 to 70 μ m, most preferably is 1 to about 50 μ m.Can be dissolved in 1.0 in the salt solution arbitrarily to the 5%W/V active ingredient solution by for example intravenous injection, or oral, for example, containing about 0.5 tablet to about 100mg/Kg activeconstituents, capsule or syrupy mode are finished.By continuous infusion provide about 0.01 to about 5.0mg/Kg/hour or at interval infusion contain to have an appointment and 0.4 keep desirable blood levels to the mode of about 15mg/Kg activeconstituents.

Though activeconstituents can be individually dosed, preferably provides with medicine type.Preparation of the present invention comprise at least a above-mentioned activeconstituents and its one or more can be by the carrier that is subjected to and any one or multiple other therapeutical agent.Various carriers must be " acceptable ", just can be compatible with other composition of preparation and to not injury of patient.

Preparation of the present invention comprises that those are fit to any aforementioned administration, and can provide with unit dosage form easily, and can be by the preparation of the known any method preparation of pharmaceutical field.These preparation methods comprise activeconstituents and the carrier blended step that constitutes one or more ancillary components.Usually, the preparation of preparation is by evenly combining closely activeconstituents and liquid vehicle or subdivided solids carrier or both, then, if desired, with product shaping.

The preparation of the present invention that is suitable for oral administration can discrete units provide, and contains the capsule of preset value activeconstituents, cachet or tablet as constituent parts; Provide with powder or particle form; Provide with solution or the form of suspension that is in moisture or the on-aqueous liquid; Or provide with liquid milk sap of oil-in-water-type or the liquid form of emulsion of water-in-oil-type.Activeconstituents also can bulk, electuary, or paste or be included in the intravital form of lipid and provide.

By will be arbitrarily one or more ancillary components together suppress or mold pressing prepares tablet.The preparation of machine tablet agent is to utilize the active compound of suitable machine stranglehold liquid form such as powder or particle, and at random with tackiness agent (for example, polyvinyl pyrrolidone, gelatin, Vltra tears), lubricant, inert diluent, sanitas, disintegrating agent (for example, primojel, crosslinked polyvinyl pyrrolidone, or tensio-active agent or dispersant croscarmellose sodium).The press back tablet can be prepared by the mixture of the moistening powder compound of inertia liquid diluent by utilizing the suitable machine mold pressing.Tablet can at random coated or score, and can be formulated to can be slowly or the controlled release form of activeconstituents wherein, can utilize as Vltra tears to change ratio, and obtain ideal release profile figure, maybe when adding liquid, for soluble or blistered.Tablet can drape over one's shoulders casing arbitrarily, thereby is provided at the intestines position rather than discharges in stomach.

Be suitable for that oral preparation also can comprise buffer reagent so as in and hydrochloric acid in gastric juice.These buffer reagents are optional from a large amount of organic or inorganic things, cry out as weak acid or with their conjugation salt blended.

Capsule can pour into vegetables pine or compressing powder by utilizing suitable filling machine, and contains one or more additives arbitrarily.Suitable additive comprises tackiness agent such as polyvinyl pyrrolidone; Gelatin, lubricant, inert diluent and the disintegrating agent described in tablet.Capsule also can or contain little pill or dispersive subsection form by preparation, so that slow or controlled release activeconstituents can be provided.Its preparation can press the mixture of wet medicine and extruding subsidiary (as Microcrystalline Cellulose) and thinner such as lactose to finish by extruding and group.The spherule that produces produces lasting release characteristics with semi-permeable membranes (as ethyl cellulose, Eudragit WE30D) coating like this.

Edible foam or foaming preparations should comprise ideally: the 50-70% edible oil, especially vegetables oil comprises Semen Maydis oil, peanut oil, Trisun Oil R 80, sweet oil and soya-bean oil, 2-10% one or more tensio-active agents, especially Yelkin TTS, polyvalent alcohol, poly-hydroxy polymerization ester comprises glycerin fatty acid ester, polyglycerol ester fatty acid ester (as four oleic acid+glyceryl ester), or sorbitan fatty acid esters (as dehydration sorbose-hard acid ester); The propelling agent that 1-4% is suitable for absorbing mainly is a compressed gas propellant, and feature is an ammonia, nitrous oxide or carbonic acid gas, or hydrocarbon gas, particularly propane, butane or Trimethylmethane; One or more particle diameters of 0.5-30% viscosity modifier in the 10-50 micrometer range, particularly powdered sucrose or colloidal silica, and can select to contain one or more suitable non-toxicity tinting material, seasonings or sweeteners of 0.5-10%.Activeconstituents is preferably in this preparation with 10-46%, and best 30% concentration exists.Aforesaid suitable food and foam or foaming preparations can prepare according to a conventional method, for example by mixing edible oil, tensio-active agent and any other soluble components add viscosity modifier, and mill grinds mixture, form the homodisperse liquid suspension.In the activeconstituents mixing row ground mixture, to disperseing fully.At last, the propelling agent with metered amount after in described mixture is adjusted to suitable decollator is incorporated in the mixture.

But the present invention is used for the surface becomes ointment, emulsifiable paste, suspension, lotion, pulvis, solution, paste, gel, sprays, aerosol or finish for the medicament preparation of usefulness.Perhaps, preparation can comprise dressing as the bandage or the surgical adhesive that soak into active ingredient and can select to contain one or more vehicle or thinner.

Dispersible tablet (discrete Patches) form that the composition that is suitable for percutaneous dosing can be suitable for keeping for a long time closely contacting with the cloudy testis of receptor is provided.These sheets are fit to contain: 1) in any aqueous buffer solution 2) be dissolved in the tackiness agent or 3) be dispersed in the active compound in the polymkeric substance.The suitable concentration of active compound is about 1% to 20%, is preferably about 3% to 15%.As a kind of specific possibility, by pressing PharmaceuticalResearch, the iontophoresis of being summarized in 3 (6), 318 (1986) spins off active compound from sheet.

For the infection of eyes or other outward sense such as mouth and skin, preferably use surperficial ointment or cream formulation, wherein contain example and go into 0.075 to 20%W/W, be preferably 0.2 to 15% W/W, most preferably be the active ingredient of 0.5 to 10%W/W amount.When preparation during ointment, activeconstituents can together use with the miscible emulsus alkali of paraffinic or water.Perhaps, activeconstituents and oil-in-water-type emulsifiable paste base or water-in-oil-type oil base together are mixed with emulsifiable paste.

If desired, the water of emulsifiable paste base can comprise, for example, 40-45%W/W polyvalent alcohol at least promptly contains the alcohol such as the propylene glycol of two or more hydroxyls, butane-1,3-glycol, mannitol, Sorbitol Powder, glycerol and polyoxyethylene glycol and their mixture thereof.Surface agent can comprise strengthening and absorbs or promote the activeconstituents compound interior or other position of working that penetrates to the skin.This class dermal osmosis accelerator example comprises methyl-sulphoxide and relevant analogue.

The oil phase of emulsion of the present invention can comprise small amounts of emulsifiers (perhaps being called Emulgents), but preferably comprise at least a emulsifying agent with fat or oil or fat and oil the two at interior mixture.As preferably, hydrophilic emulsion comprises the oleophylic emulsion as stablizer simultaneously.Preferably comprise oil ﹠ fat simultaneously.Emulsion and or do not have stablizer together to form so-called emulsifying wax, this wax and oil and/or fat together form so-called emulsification ointment base, thereby have formed the oil phase in the ointment formulation.

The emulsifying agent and the emulsion stablizer that are fit to preparation use of the present invention comprise polysorbate60, sorbester p17, cetostearyl alcohol, tetradecyl alcohol, glycerine list hard acid ester and sodium lauryl sulphate.

The mountain is lower in the solubleness of the oil that may be used for the pharmaceutical emulsion preparation in a large number in active compound, so be suitable for the oil of preparation or fatty selection according to the cosmetic result that requires to obtain.Emulsifiable paste should be preferably non-grease, and is pollution-free and can wash product, and should have suitable consistency, avoids also leaking the container from pipe or its.Can use straight or branched, monobasic or binary alkyl ester are as two dissidents, two acid esters, iso-spermaceti ester alcohol stearic acid, the propylene glycol ester of coconut fatty acid, 14 (alkane) isopropyl propionate, decyl oleate, Wickenol 111, butyl stearate, 2-ethylhexyl cetylate or branched ester mixture are Crodamol CAP, back three kinds of esters are preferred ester.These esters can use separately or be used in combination according to needed character.On the other hand, also can use high melting-point lipid such as white soft wax and/or whiteruss or other Dormant oils oil.

The preparation that is suitable for the eyes topical also comprises eye drops, and wherein activeconstituents is to be dissolved or suspended in the suitable carriers, especially water-containing solvent.In this preparation, the concentration of activeconstituents is preferably 0.5 to 20%, is 0.5 to 10% more preferably, is in particular about 1.5%W/W.

The preparation that is suitable for oral cavity local medication comprises that activeconstituents is dispersed in the flavoring substance, be generally sucrose and gum arabic or the yellow lozenge that holds in the glue, activeconstituents is included in inert material such as gelatin, or the pastille within sucrose and the gum arabic, and activeconstituents is included in the washing liquid of oral cavity within the suitable liquid carrier.

The rectal administration preparation can suppository form provide, and the suitable base (Suitable base) that comprises as theobroma oil or high fatty alcohol (as hard wax, European Pharmacopoeia) or triglyceride level and saturated fatty acid (as Witepsol) is arranged

Be suitable for intranasal administration wherein carrier be the meal of 20 to 500 micrometer ranges for example for the solid preparation comprises particle size, meal is pressed the mode that snuff uses and is given usefulness, promptly is drawn in the nasal passage rapidly from being installed near in the powder container of nose.Be suitable for nasal spray or the administration of nose drops mode wherein carrier comprise the water or the oil solution of activeconstituents for the fluidic preparation.

The preparation that is suitable for vagina administration also can be a vaginal suppository, stopper, and emulsifiable paste, gel, paste, foaming agent or sprays wherein also comprise this professional known suitable carrier except that activeconstituents.

The preparation that is suitable for parenterai administration comprises moisture and non-water aseptic parenteral solution, wherein can close antioxidant, buffer reagent, bacteriostatic agent and the solute that the component isotonicity can be provided to predetermined receptor's blood, and moisture and non-water sterile suspension, wherein can comprise suspension agent and thickening material.Preparation can unitary dose or multi-dose container for example the form of ampoule and phial provide, and can under freeze-drying (lyophilize) condition, preserve, before the use, only need directly adding sterile liquid carrier, for example water for injection.Interim injection solution and suspension can aforesaid sterile powders, and granula and tablet make.

Preferred unit dose formulations contains per daily dose or divided dose (as previously mentioned), or its an amount of part activeconstituents

Should be appreciated that the composition of mentioning except the front is specific that preparation of the present invention also can comprise the therapeutical agent that described this specialty of preparation type is habitual, for example, also can comprise seasonings for the preparation of oral administration.

The present invention further comprises the salt of preparation formula (I) compound or formula (I) compound, ester or have the method for derivative or any its solvate of physiological function, comprise following any:

(A) with the purine bases of formula (II) Wherein R ' definition or is its sense equivalent as above, with formula (III) compound reaction production (I) compound; R wherein ⁴Represent hydrogen or hydroxyl protecting group, A represents bound phosphate groups or its salt or purine except that (II) or pyrimidine part, or represents leavings group; Or

(B) make formula (IV) compound Wherein the definition of R ' is the same, and Z represents the precursor group of fluorine atom, with reagent and/or in making the erythro form configuration precursor group Z convert under the condition of fluorine atom and react; And then, or meanwhile, carry out a following step or multistep and transform arbitrarily:

(i) remove any remaining protecting group:

(ii) after formula (I) compound generates, be converted into formula (I) compound

Salt, ester or derivative with physiological function; Or

(iii) when the salt of formula (I) compound, ester or have the derivative of physiological function or after any its solvate generates, with this derivative conversion accepted way of doing sth (I) compound, or the different derivatives of a conversion accepted way of doing sth (I) compound.

In above-mentioned the inventive method, formula (II), starting compound (III) and (IV), and mentioned reagent and condition can be selected the known relevant art of nucleosides synthetic chemistry specialty for use.The example of this class method for transformation directiveness is as follows told, and should be known in according to desired formula (I) compound and improve ordinary method.Especially when described conversion might cause the undesirable reaction of unstable group in addition, then this group should be protected with ordinary method, removed protecting group subsequently after converting.

According to carrying out the used condition of method A, formula (II) purine bases and formula (III) compound can use maybe needn't use known protecting group protection, these protecting groups such as acyl group; for example; alkanoyl (for example ethanoyl), the alkanoyl of replacement is as the oxyalkyl chain alkyloyl; aroyl (as benzoyl); ether, for example, trialkylsilkl; as t-butyldimethylsilyl or other group, as aralkyl (for example benzyl) or phosphate-based.

This class group can utilize acid or basic hydrolysis, hydrogenolysis, or the enzyme catalysis mode is removed.Acyl group is general with crying out hydrolysis method to remove, and silyl is removed with acid hydrolysis or fluorion.The most handy catalytic hydrogenolysis mode of aralkyl such as benzyl is removed.

Usually there are two kinds of methods to be used to carry out method A, i.e. enzyme catalysis and chemical process.

Method (A) can be by for example making suitable formula (II) purine bases, and wherein the definition of R ' as above or is represented any its sense equivalent, and for example salt or protected derivative (on seeing) is finished with being reflected under the enzyme catalysis of formula (III) compound, wherein R ⁴Represent hydrogen or hydroxyl protecting group (on seeing), A represents purine or pyrimidine part (except that (II)), phosphate-based or its salt.

When A represents purine or pyrimidine derivatives (except that (II)), reaction can be carried out in the presence of following enzyme; (i) Starch phosphorylase, as purine nucleoside phosphorylase and thymidine phosphorylase and be in organophosphate or its salt in Starch phosphorylase, or (ii) transferring enzyme, for example, N-ribodesose based transferase.In order to obtain The compounds of this invention, when this method of use, formula (III) compound should be beta configuration.

According to Kren itsky etc., Biochemistry, 20,3615-, 1981 and US4, method described in 381,344 prepares fast nucleoside phosphorylase and the thymidine phosphorylase of making.

N-ribodesose based transferase can separate from following bacterial strain according to the routine biochemistry technology and obtains:

(I) colon bacillus strain SS6030/14, its expression Bacterium lacticum enzyme is from American

Type Culture Collection (ATCC) Rockville obtains,

MD20852-1776 on July 18 nineteen ninety, steps on

The record number: ATCC68367, or

(ii) colon bacillus strain SS70-8/15 represents the Bacterium lacticum enzyme, from

ATCC obtains, on June 17th, 1992, accession number: ATC

C69016。

When A represented phosphate-based or its salt, reaction can be at single Starch phosphorylase, and the platform purine nucleoside phosphorylase carries out under existing.For obtaining The compounds of this invention, when using present method, formula (III) compound should be α-configuration.

Can use blocking group in the enzymatic process, but find in the actually operating there is no need, and in some cases, in fact unfavorable to overall yield.

Method (A) can be by for example making definition formula (II) compound as above and the reaction of formula (III) compound, wherein R ⁴Represent hydrogen or hydroxyl protecting group, and the suitable leavings group of A representative, resemble halogen atom, as chlorine; Acyloxy, as acetoxyl group or alkoxyl group, as methoxyl group, in the presence of catalyzer, as tin chloride (IV) or trimethylammonium fluoroform sulphonate, chemical catalysis carries out and finishes in suitable solvent such as acetonitrile.

With enzymatic process relatively, found in the chemical catalysis method (a) formula (II) and (III) compound preferably protected (referring to above), and (b) formed formula (I) compound is α-and the mixture of β-anomers.β-anomers of the present invention can obtain as silica gel column chromatography or HPLC method separation anomers by facile method in known method of professional and technical personnel or the chemical literature.

R wherein ¹Definition as above, or represent formula (II) compound of any its sense equivalent to obtain from market, for example buy from Aldrich Chemical Company, or, for example, use and the identical or similar approach described in the following document: Robins etc. by facile method preparation in known method of professional or the chemical literature, J.Amer, Chem.Soc., 1957,79,490-494 and Montgomery and Temple, J.Amer.Chem.Soc., 1961,83,630-635.

For example, suitable purine bases can be suitable leavings group by 6-substituting group wherein, prepares by group as described in the nucleophilic displacement as the corresponding purine of chlorine.Thereby wherein the 6-substituting group is that the purine of propoxy-formula cyclobutoxy group can be by in the presence of alkali such as sodium hydride, handle corresponding 6-chloropurine preparation with corresponding propyl alcohol or cyclobutanol, and wherein the 6-substituting group is a cyclopropylamino, piperidino-(1-position only), or the purine of pyrrolidyl can be by in suitable solvent, with suitable amine, and the cyclopropylamine of promptly respectively doing for oneself, piperidines, or tetramethyleneimine is handled corresponding 6-chloropurine and is prepared.

2-amino-6-chloropurine precursor can obtain (Aldrich ChemicalCo) from the market, or by facile method preparation in known method of professional or the chemical literature.

Wherein on behalf of formula (III) compound of pyrimidine or purine part, A be easy to by facile method preparation in known method of professional or the chemical literature.For example, 2 ' 3 '-dideoxy-3 '-floxuridine can obtain from the market or by A.Kowollick etc. at J.Prakt.Chem., 1973,315 (5), method described in 895 preparation, 3 '-deoxidation-3 '-fluorothymidine can be by Etzo l d etc. at Tetrahedron, 1971,27, the preparation of method described in the 2463-2472, and 2 ', 3 '-dideoxy-3 '-the fluorine guanine can be by Herdewijn etc. at J.Med.Chem., 1988,31, the preparation of method described in the 2040-2048.

Wherein phosphate-based formula (III) compound of A representative can be by being similar to the method chemical catalysis preparation described in the chemical literature, or to represent formula (III) compound of pyrimidine or purine part by A wherein be raw material, handles with Starch phosphorylase such as thymidine phosphorylase to prepare.

R wherein ⁴As defined above, and on behalf of formula (III) compound of leavings group such as methoxyl group, A can obtain from the market, or press Asahi Glass and prepare in method described in the Japanese patent application No. JP0129390.

As for method (B), can on behalf of formula (IV) compound of leavings group, Z finish by for example handling wherein with suitable fluorizating agent, wherein said fluorizating agent such as diethylaminosulfurtrifluoride, Potassium monofluoride, potassium hydro-fluoride, or the tetra-n-butyl Neutral ammonium fluoride, described leavings group such as hydroxyl or organic sulfonyloxy, weevil sulfonyloxy or trifluoro-methanesulfonyl oxy.

Formula (IV) compound can be by facile method preparation in known method of professional or the chemical literature, for example, uses Herdewijn etc. at J.Med-Chem., and 1988,31, method described in the 2040-2048.

Ester of the present invention can for example, be handled formula (I) compound parent with suitable esterifying agent by this professional currently known methods preparation, handles as resemble muriate or acid anhydrides with suitable acyl halide.

Formula (I) compound can change into the physiologically acceptable ether of corresponding formula (I) compound by currently known methods and suitable alkylation reactions.

Formula (I) compound comprises its ester, can be by currently known methods as change into physiologically acceptable salt with suitable alkaline purification.The ester of formula (I) compound or salt also can change into parent compound by the mode as hydrolysis.

Following embodiment only is used to illustrate the present invention, and must not think the restriction scope of the invention.The salt of predicate ' activeconstituents ' expression (I) compound that embodiment is used or formula (I) compound, ester or derivative, or any its solvate with physiological function.Embodiment 12-amino-9-(2, the 3-dideoxy-red moss furan pentose of 3-fluoro-beta-D-base)-6-positive propoxy-9H-purine

Utilize propyl alcohol (Aldrich Chemical Company) to replace in the presence of sodium hydride that the chlorine atom prepares 2-amino-6-positive propoxy-9H-purine in 2-amino-6-chloropurine (Aldrich Chemical Company).With 2-amino-6-positive propoxy-9H-purine (0.50g, 2.6mmoles) be dissolved in dimethyl formamide (5ml, DMF) and methyl-sulphoxide in (5ml, DMSO).Add 3 '-deoxidation-3 '-fluorothymidine (0.76g, 3.1mmoles) and 30ml contain the 10mM potassium phosphate buffer (pH6.8) of 0.04% potassium azide.In reaction mixture, add the purifying purine nucleoside phosphorylase be adsorbed on the 5ml diethylaminoethyl cellulose resin (7,500I.U.) and thymidine phosphorylase (3750I.U.), and with suspension 37 ℃ of stirrings.After 24 days, filtering reaction, filtrate puts in a succession of multistage column.First post is equipped with AG1-X2 (OH-type, 2.5 * 10cm), and second post is equipped with the Amberlite XAD-2 resin.After sample adds, post water (500ml) washing, product methanol-eluted fractions.(4.8 * 20cm) purification by flash chromatography, eluent are methylene dichloride through silicagel column will to contain the part of product then; Methyl alcohol (95: 5).Vacuum evaporating solvent, lyophilize obtains 0.46g 2-amino-9-(2, the 3-dideoxy-red moss furan pentose of 3-fluoro-beta-D-base)-6-propoxy--9H-purine (50%); Mp128 ℃; TLC Rf0.85 (silica gel, MeCN; 15N NH ₄OH: H ₂O/85: 5: 10); [α] _D-37.2 (C=0.5, DMF); UV λ max (∈ * 10 ^-3) when pH 7; 280 (9.9) and 247.5 (10.3); When pH 13; 279.5 (10.6) and 247.5 (10.6); ¹H NMR (200MHz, DMSO-d ₆) δ 8.07 (s, 1H, H ₈), 6.41 (b, 2H, NH ₂), 6.23 (dd, 1H, H ₁,, J=9.1Hz, J=5.7Hz), 5.37 (dd, 1H, H ₃,, J=53.8Hz, J=4.2Hz), 5.19 (t, 1H, OH _{5 '}, J=5.6Hz), 4.34 (t, 2H, J=6.8Hz, OCH ₂CH ₂CH ₃), 4.16 (dt, 1H, H, J=27.0Hz, J=4.9Hz), 3.56 (m, 2H, H ₅' and H _{5 "}), 2.94 and 2.68 (2m, total 2H, H _{2 '}And H _{2 "}), 1.73 (m, 2H, OCH ₂CH ₂CH ₃) and 0.95 (t, 3H, J=7.4Hz, OCH ₂CH ₂CH ₃): MS (Ci) 312 (M+1), 293 (M-F), 194 (MH ₂-C ₅H ₈FO ₂).Analyze (C ₁₃H ₁₈FN ₅O ₃0.16H ₂O) C, H, F, N; Calculated value; C, 49.70; H, 5.88; F, 6.05; N, 22.29 measured values; C, 49.58; H, 5.90; F6.30; N22.04.Embodiment 22-amino-6-cyclobutoxy group-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine

Utilize cyclobutanol (Aldrich Chemical Company) to replace in the presence of sodium hydride that the chlorine atom prepares 2-amino-6-cyclobutoxy group-9H-purine in 2-amino-6-chloropurine (Aldrich Chemical Company).

With 2-amino-6-cyclobutoxy group-9H-purine (0.50g, 2.4mmoles) and 2 ', 3 '-dideoxy-3 '-floxuridine (0.67g, 2.9mmoles) be suspended in the potassium phosphate buffer that contains 0.04% potassium azide (50ml, 10mM, pH7.0).In reaction mixture, add the purine nucleoside phosphorylase (1120.I.U.) be fixed on the real resin of diethyllaminoethyl fiber and thymidine phosphorylase (10,000I.U.), and with suspension in 45 ℃ of stirrings down.After 8 days, in reactant, add methyl alcohol (150ml).Reactant is applied to and is loaded with AG1-X2 (OH ^-Type in 2.5 * 10cm) the post, is used methanol-eluted fractions.Compile the part that contains product, concentrate, and (2.5 * 20cm) chromatogram purifications, eluent are methylene dichloride: acetone (92: 8) through quick silica gel.Remove and desolvate and lyophilize, obtain 0.57g 2-amino-6-cyclobutoxy group-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine (70%): mP139-140 ℃; TLC Rf0.75 (silica gel, MeCN: 15N NH ₄OH: H ₂O/85: 5: 10); [α] _D ²⁰-17.2 ℃ (C=0.5, DMF); UV λ max (∈ * 10 ^-3) when pH7; 281 (9.5) and 247 (9.4): when pH13,281.5 (9.9) and 247 (9.5); ¹H NMR (400MHz, DMSO-d ₆) δ 8.08 (s, 1H, H ₈), 6.40 (s, 2H, NH ₂), 6.22 (dd, 1H, H _{1 '}, J=9.4Hz, J=5.5Hz), 5.38 (dd, 1H, H _{3 '}, J=53.6Hz, J=4.0Hz), 5.26-5.30 (m, 1H, OCH), 5.19 (t, 1H, OH _{5 '}, J=5.6Hz), 4.16 (dt, 1H, H _{4 '}, J=26.8Hz, J=4.8Hz), 3.55 (t, 2H, H _{5 '}And H _{5 "}, J=5.3Hz), 2.78-2.96 and 2.52-2.64 (m, 2H, H _{2 '}With _{2 "}) and 2.36-2.45,2.05-2.16,1.74-1.83, and 1.55-1.67 (4m, 6H, cyclobutyl proton); MS (ci) 324 (M+1), 304 (M-F), 206 (MH ₂-C ₅H ₈FO ₂). ultimate analysis C ₁₄H ₁₈FN ₅O ₃0.65H ₂O calculated value: C, 50.19; H, 5.81; F, 5.67; N, 20.90 measured values: C, 50.27; H, 5.74; F, 5.62; N, 20.77 embodiment 32-amino-6-(cyclopropylamino)-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine

Utilize the chlorine atom in cyclopropylamine (Aldrich Chemical Company) displacement 2-amino-6-chloropurine (Aldrich Chemical Company) to prepare 2-amino-6-(cyclopropylamino)-9H-purine.

With 2-amino-6-(cyclopropylamino)-9H-purine (0.50g; 2.7mmoles) be dissolved among DMF (5ml) and the DMSO (5ml), add 3 '-deoxidation-3 '-fluorothymidine (0.81g, 3.3mmoles) and contain 0.04% potassium azide potassium phosphate buffer (10mM, 30ml, pH6.8).In reactant, add purine nucleoside phosphorylase (7500I.U.) and the thymidine phosphorylase (37.50I.U.) that is fixed on the diethylaminoethyl cellulose resin, and suspension is stirred down at 37 ℃.After 23 days, will react filtration, filtrate is applied in a succession of multistage column.Be loaded with AG1-X2 (OH-type, 2.5 * 10cm), and be loaded with Amberlite XAD-2 resin (2.5 * 20cm) in second post in first post.After sample adds, post water (700ml) washing, product methanol-eluted fractions.(4.8 * 24cm) purification by flash chromatography, eluent are ethyl acetate: methyl alcohol (95: 5) through silicagel column to contain the part of product then.Further (2.5 * 40cm) purification by flash chromatography, eluent are methylene dichloride to product: methyl alcohol (9: 1) through silicagel column.Vacuum evaporating solvent and lyophilize obtain 0.30g 2-amino-6-(cyclopropylamino)-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine (33%): mp75 ℃ of TLC Rf0.66 (silica gel, MeCN: 15N NH ₄OH:H ₂O/85: 5: 10); [α] _D-26.4 (C=0.5, DMF); UV λ max (∈ * 10 ^-3) when pH7; 283 (13.7) and 261 (3.2); When pH13,283 (13.8) and 261 (9.3); ¹H NMR (300MHz, DMSO-d ₆) δ 7.93 (s, 1H, H ₈), 7.44 (d, 1H, NH, J-3.8Hz), 6.22 (dd, 1H, H _{1 '}, J=9.4Hz, J=5.6Hz), 5.85 (b, 2H, NH ₂), 5.52 (t, 1H, OH _{5 '}, J=5.9Hz), 5.40 (dd, 1H, H _{3 '}, J=54.1Hz, J=4.4Hz), 4.19 (dt, 1H, H _{4 '}J=27.5Hz, J=4.3Hz), 3.60 (apparent triplet, 2H, H _{5 '}And H _{5 "}, J=5.1Hz), 2.97,2.81 and 2.58 (3m, 3 H altogether, H _{2 '}, H _{2 "}And-CH ₂CH ₂CH) and 0.66 and 0.58 (2m, 4H, CH ₂CH ₂CH); MS (ci) 309 (M+1), 289 (M-F), 191 (MH ₂-C ₅H ₈F-O ₂). analyze (C ₁₃H ₁₇FN ₆O ₂, 0.6H ₂O0.3C ₂H ₆O) C, H, F, N: calculated value: C, 49.06; H, 6.05; F, 5.71; N25.24 measured value: C, 49.06; H, 5.84; F, 5.97; N25.00 embodiment 42-amino-6-(1-piperidino-(1-position only))-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine

Utilize the chlorine atom in piperidines (Aldrich Chemical Company) displacement 2-amino-6-chloropurine (Aldrich Chemical Company) to prepare 2-amino-6-(1-piperidino-(1-position only))-9H-purine.

With 2-amino-6-(1-piperidino-(1-position only))-9H-purine (0.60g, 2.7mmoles) and 2 ', 3 '-dideoxy-3 '-floxuridine (0.50g, 2.2mmoles) be suspended in the potassium phosphate buffer that contains 0.04% potassium azide (50ml, 10mM, pH7.0).In reactant, add the purine nucleoside phosphorylase (1120I.U.) be fixed on the diethylaminoethyl cellulose and thymidine phosphorylase (10,000I.U.) (Krenitsky, etal., Biochemistry, 20,3615,1981 and US patent 4,381,344), and at 45 ℃ of stirred suspensions.After 3 days, in reactant, add methyl alcohol (100ml).Reactant is applied to and is loaded with AG1-X2 (the OH-type in 2.5 * 10cm) posts, and is used the MeOH wash-out.Collection contains the product part, concentrate, and (2.5 * 20cm) purification by flash chromatography, eluent is methylene dichloride: acetone (9: 1) through silica gel.Remove and desolvate and lyophilize, obtain 0.41g 2-amino-6-(1-piperidino-(1-position only))-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine (55%): mp101-104 ℃; [α] _D ²⁰-15.6 (C=0.50, DMF); UV λ max (∈ * 10 ^-3) when pH7; 287 (17.6); When pH13,287 (17.6); ¹H NMR (300MHz, DMSO-d ₆) δ 7.97 (s, 1H, H ₈), 6.26 (dd, 1H, H _{1 '}, J=9.4Hz, J=5.6Hz), 5.86 (b, 2H, NH ₂), 5.50 (t, 1H, OH _{5 '}, J=6.0Hz), 5.40 (m, 1H, H _{3 '}), 4.25 and 4.14 (2m, 5H, H _{4 '}, CH ₂-N-CH ₂), 3.60 (t, 2H, H _{5 '}And H _{5 "}, J=4.9Hz), 2.89 and 2.60 (2m, 2H, H _{2 '}With _{2 "}), 1.67 and 1.55 (2m, 6H, (CH ₂) ₃); MS (ci) 337 (M+1), 317 (M-F), 219 (MH ₂-C ₅H ₈FO ₂). ultimate analysis C ₁₅H ₂₁FN ₆O ₂0.35H ₂O: calculated value: C, 52.58; H, 6.38; F, 5.54; N, 24.52 measured values: C, 52.78; H, 6.50; F, 5.42; N, 24.38 embodiment 52-amino-6-(1-pyrrolidyl)-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine

Utilize the middle chlorine atom of tetramethyleneimine (Aldrich Chemical Company) displacement 2-amino-6-chloropurine (Aldrich Chemical Company) to prepare 2-amino-6-(1-pyrrolidyl)-9H-purine.

With 2-amino-6-(1-pyrrolidyl)-9H-purine (0.59g, 2.8mmoles) and 2 ', 3 '-dideoxy-3 '-floxuridine (0.50g, 2.2mmoles) be suspended in the potassium phosphate buffer that contains 0.04% folded ammonification potassium (50ml, 10mM, pH7.0).In reactant, add the purine nucleoside phosphorylase (1120I.U.) be fixed on the diethylaminoethyl cellulose and thymidine phosphorylase (10,000I.U.) (Krenitsky, etal., Biochemistry, 20,3615,1981 and US patent 4,381,344), and at 45 ℃ of stirred suspensions.After 3 days, in reactant, add methyl alcohol (10ml).During reactant is executed to being loaded with AG1-X2 (OH ^-Type in 2.5 * 10cm) the post, and is used the MeOH wash-out.Collection contains the product part, concentrate, and (2.5 * 20cm) purification by flash chromatography, eluent is methylene dichloride: methyl alcohol (97: 3) through silica gel.Remove and desolvate and lyophilize, obtain 0.491g 2-amino-6-(1-pyrrolidyl)-9-(2, the red moss of 3-dideoxy-3-fluoro-beta-D--furan pentose base)-9H-purine (6 8%): mp101-104 ℃; [α] _D ²⁰-15.2 (C=0.5, DMF); UV λ max (∈ * 10 ^-3) when pH7; 283 (16.0); With 267 (11.4) (sh); When pH13,284 (16.0); With 266 (11.2) (sh); ¹H NMR (300MHz, DMSO-d ₆) δ 7.94 (s, 1H, H ₈), 6.26 (dd, 1H, H _{1 '}, J=9.4Hz, J=5.6Hz), 5.82 (b, 2H, NH ₂), 5.53 (t, 1H, OH _{5 '}, J=5.6Hz), 5.42 (dm, 1H, H _{3 '}, J=53.5Hz), 4.20 (dm, 1H, H _{4 '}, J=27.3Hz), 3.96 (b, 4H, 2CH ₂), 3.60 (t, 2H, H _{5 '}And H _{5 "}, J=4.9Hz), 2.90 and 2.55 (m, 2H, H _{2 '}And H _{2 "}) and 1.91 (b, 4H, (CH ₂) ₂); MS (cf) 323 (M+1), 303 (M-F), 205 (MH ₂-C ₅H ₈FO ₂). ultimate analysis C ₁₄H ₁₉FN ₆O ₂0.35H ₂O: calculated value: C, 51.17; H, 6.04; F, 5.78; N, 25.57 measured values: C, 51.14; H, 5.83; F, 5.58; N, 25.36 embodiment, 6 tablet formulations

Following preparation A, B and C add Magnesium Stearate subsequently by with each composition and polyvinyl pyrrolidone solution wet granulation, compressing tablet and getting.Preparation A

Every (a) activeconstituents 250 250 of every mg/ of mg/ (b) lactose B.P. 210 26 (c) polyvinyl pyrrolidone B.P. 15 9 (d) sodium starch glycollate 20 12 (e) Magnesium Stearate 53

500 300 preparation B

Every (a) activeconstituents 250 250 of every mg/ of mg/ (b) lactose 150 (c) Microcrystalline Cellulose pH101 60 26 (d) polyvinyl pyrrolidone B.P. 15 9 (e) sodium starch glycollate 20 12 (f) Magnesium Stearate 53

500 300 formulation C

Every activeconstituents of mg/ 100 lactose 200 starch 50 polyvinyl pyrrolidones 5 Magnesium Stearates 4

359

Following preparation D and E get by directly suppressing mixing element.Used lactose is direct compression (Dairy Crest-" Zeparox ") among the preparation E.Preparation D

Every activeconstituents of mg/ 250 pregelatinized Starch NF15 150

400 preparation E

Every activeconstituents of mg/ 250 lactose 150 Microcrystalline Celluloses 100

500 preparation F (sustained release dosage)

This preparation adds Magnesium Stearate subsequently by with composition (following) and polyvinyl pyrrolidone solution wet granulation, compressing tablet and getting.

Every (a) activeconstituents 500 of mg/ (b) Vltra tears 112 (Methocel K4M Premium) is lactose B.P. 53 (d) polyvinyl pyrrolidone B.P. 28 (e) Magnesium Stearate 7 (c)

700

Medicine begins to discharge in about 6-8 hour scope, and discharges fully after 12 hours.Embodiment 7 capsule preparations preparation A

Capsule preparations is by mixing each composition of preparation D in the foregoing description 2, and mixture is packed in the two joint hard gelatin capsules and gets.Preparation B (down) can prepare by similar approach.Preparation B

Mg/ capsule (a) activeconstituents 250 (b) lactose B.P. 143 (c) sodium starch glycollate 25 (d) Magnesium Stearate 2

420 formulation C

Mg/ capsule (a) activeconstituents 250 (b) Macrogol 4000 B.P. 350

600 preparation D

Mg/ capsule activeconstituents 250 Yelkin TTS 100 peanut oil 100

450

The capsule of preparation D be by the dispersed activity composition in Yelkin TTS and peanut oil, and dispersion be packed in the soft elastic gelatin capsule and get.Preparation E (slow releasing capsule)

Following slow release capsule preparation is by utilizing extrusion machine extruding composition (a), (b) and (c), and subsequently with extrudate nodularization and dry.Dry piller is so used release membranes (d) coating and is packed into two joint hard gelatin capsules and gets.

Mg/ capsule (a) activeconstituents 250 (b) Microcrystalline Cellulose 125 (c) lactose B.P. 125 (d) ethyl cellulose 13

51 embodiment, 8 injection formulations preparation A activeconstituents 0.200g0.1M hydrochloric acid solns, or the 0.1M sodium hydroxide solution in right amount to pH 4.0 to 7.0 sterilized waters in right amount to 10ml

Activeconstituents is dissolved in most of water under 35 ℃-45 ℃, regulates pH between 4.0 and 7.0 with proper concn hydrochloric acid or sodium hydroxide.Every batch of water is supplemented to volume then, filters in the aseptic 10ml amber glass bottle (type1) by aseptic millipore filtration, utilizes sterile seal and outside seal sealing by fusing.Preparation B activeconstituents 0.125 is aseptic, pyrogen-free, pH be 7 phosphate buffer in right amount to 25ml embodiment 9 intramuscularly agent activeconstituents 0.20g benzylalcohol 0.10gGlycofurol75 1.45g waters for injection in right amount to 3.00ml

Activeconstituents is dissolved in the glycofurol, adds benzylalcohol then and makes dissolving, adds water to 3ml.Mixture is by aseptic filtering with microporous membrane, the amber phial of the 3ml of sealed, sterile (type1).Embodiment 10 syrup activeconstituents 0.25g Sorbitol Solution USP 1.50g glycerine 2.00g Sodium Benzoate 0.005g seasoningss, Peach 17.42.3169 0.0125ml purified water is in right amount to 5.00ml

Activeconstituents is dissolved in the mixture of glycerine and most of purified water.In solution, add the Sodium Benzoate aqueous solution then, add Sorbitol Solution USP subsequently, add seasonings at last.Add purified water to volume, mix.0.125g is aseptic for preparation B activeconstituents, pyrogen-free, and pH is that 7 phosphate buffered saline buffer is in right amount to 25ml embodiment 11 suppositorys

Mg/ suppository activeconstituents (63 μ m) ^*250Hard Fat, B.P. (Witepsol H15-Dynamit NoBel) 1770

The used activeconstituents of 2020* is a powder, and wherein at least 90% particle diameter is 63 μ m or littler.

Witepsol H15 with 1/5th is melted in the vapour jacket pot under 45 ℃ of top temperatures.Activeconstituents is by 200 μ m screening, and is added in the molten caustic soda, utilizes the Silverson that has the track icking tool to mix, till forming level and smooth dispersion.Keep mixture in 45 ℃, in suspension, add remaining Witepsol H15, stir, mix to guarantee homogeneous phase.All suspension by 250 μ m stainless (steel) wires, makes it be chilled to 40 ℃ under constantly stirring.In 38 ℃ to 40 ℃ temperature, the 2.0g mixture is injected in the suitable 2ml plastic pattern, cooling suppository is to room temperature.Embodiment 12 vaginal suppositories

Mg/ vaginal suppository activeconstituents (63 μ m) 250 anhydroglucoses (Amhydrate Dextrose) 380 yam starchs 363 Magnesium Stearates 7

1000

Above-mentioned each composition is mixed, directly push formed mixture and just can prepare vaginal suppository.The antiviral activity of 13 pairs of hepatitis B viruses of embodiment

The anti-hepatitis B virus activity of formula (I) compound is pressed Korba B.E. and MilmanG, Antiviral Research, and 1991, Vol.15, method described in the pp217-228 is measured.

Test is used by people such as Sells at PNAS84, narration and the HePG2 that characterize in 1005 (1987) and J.Virol.62,2836 (1988), and 2.2.15 human body HBV founder cell system, and showed the hepatocellular HBV characteristic of a lot of chronic infections is all arranged.This clone proves communicable by the pathogenecity to black raw meat raw meat.This clone of external use is to determine that The compounds of this invention has anti-HBV activity.

Be the antiviral activity of test compounds, handled the monolayer cell substratum 9 days with compound (50-200 μ m).The upper strata of taking to contain extracellular virus body DNA (Dane particle) respectively at the 0th, the 4 and 9 day liquid culture medium that disappears, with Proteinase K (1mg/ml) and sodium lauryl sulphate (1%) processing, and 50 ℃ of incubations one hour.DNA uses chloroform extraction then with equal-volume phenol earlier, then with ammonium acetate and propyl alcohol precipitation.Utilize SchleiCher and Schuell method (S﹠amp; S, 10 Optical AVe., Keene, NH, 03431, Publication700,1987) with the DNA resolution of precipitate and collect on the soluble cotton, and press Southern at J.Mol.Biol.98, method described in 503 (1975) is handled.Take cell, cell obtains DNA in the cell after dissolving with the isothiocyanic acid guanidinesalt.DNA is by handling with the same procedure of handling extracellular DNA in the cell.With ammonium acetate and propyl alcohol post precipitation, with DNA resolution of precipitate in the cell,, press the described method of Southern then and handle with the restriction endonuclease HindIII cutting that is attached on the agaropectin, determine reproducible intermediate quantity.The antiviral effect of medicine is determined by the 100 demultiplications similar minimizing few and time multiplexed cell system intermediate that measurement is squeezed in the Dane particle number in the substratum at least.

Extracellular HBV DNA

(pg/ml substratum)

Day0 Day4 Day9 untreated cell 75 98 74

70 49 73

73 110 78

72 110 95 handle cell embodiment 1 (100 μ m) 35 10 1

83 7 0

77 5 0

75 16 13 (100μm) 59 13 0

58 47 1

55 36 0.1

76 39 04 (24.8μm) 86 63 7

53 41 1

74 75 6

65 59 65 (24.3μm) 95 43 14

91 30 8

86 49 3

100 50 10

Claims (7)

1. the salt of formula (I) compound or its physiologically acceptable salt, ester or this ester:

R wherein ¹Represent cyclopropyl amino or 1-pyrrolidyl.

2. according to the compound of claim 1, it is 2-amino-6-cyclopropyl amino-9-(2, the 3-dideoxy-red moss furan pentose of 3-fluoro-beta-D-base)-9H-purine.

3. according to the compound of claim 1, the salt of wherein said its physiologically acceptable salt, ester or this ester be selected from carboxylicesters, sulphonate, amino acid ester, list-, two-or three-phosphoric acid ester and salt.

4. one kind is used for the treatment of or the pharmaceutical composition of prophylaxis of viral infections, and it comprises the compound of claim 1-3 and suitable pharmaceutically useful carrier thereof.

5. according to the pharmaceutical composition of claim 4, it is tablet or capsule.

Among the claim 1-3 each compound preparation be used for the treatment of or the medicine of prophylaxis of viral infections in application.

7. prepare the method for formula (I) compound of claim 1 or 2, comprising:

(A) make formula (II) purine bases R wherein ¹Definition with described in the claim 1, or be its sense equivalent, with the reaction of formula (III) compound, production (I) compound

R wherein ⁴Represent hydrogen or hydroxyl protecting group, A represents phosphate-based or its salt, or purine except that (II) or pyrimidine part, or leavings group; Or

(B) make formula (IV) compound

R wherein ¹Definition with described in the claim 1, Z represents the precursor group of fluorine atom; With can make in the erythro form configuration precursor group Z convert the reagent of fluorine atom to and/or under such condition, react; And then or meanwhile, carry out a following step or multistep and transform arbitrarily:

(i) remove any residual protecting group;

(ii) after formula (I) compound generates, be converted into salt, the ester of formula (I) compound or derivative with physiological function; Or

(iii) when salt, the ester of formula (I) compound or have the derivative of physiological function or after any its solvate forms, with this derivative conversion accepted way of doing sth (I) compound, or the different derivatives of a conversion accepted way of doing sth (I) compound.